LOCUS       sequence-94445-e         832 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses 
            tagRFP via P2A cleavage site. EFS Promoter driven.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 832)
  AUTHORS   Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey 
            ME, Thielke A, Aster JC, Regev A, Ebert BL
  TITLE     Generation of mouse models of myeloid malignancy with combinatorial 
            genetic lesions using CRISPR-Cas9 genome editing.
  JOURNAL   Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951.
  PUBMED    24952903
REFERENCE   2  (bases 1 to 832)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951."
COMMENT     Sequence Label: pL-CRISPR.EFS.tRFP
FEATURES             Location/Qualifiers
     source          1..832
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     126..145
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        222..297
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    345..462
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        531..742
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      762..771
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             771..794
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             801..821
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
       61 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      121 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      181 tcttgtggaa aggacgaaac accggagacg tgaccgtctc tgttttagag ctagaaatag
      241 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
      301 tttgaattct agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
      361 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
      421 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
      481 agcagagatc cactttggcg ccggctcgag tggctccggt gcccgtcagt gggcagagcg
      541 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      601 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      661 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      721 cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg gattacaaag
      781 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac gg
//
